KR100943839B1 - Method for the production of bio-imaging nanoparticles with high yield by early introduction of irregular structure - Google Patents

Abstract

The present invention relates to a method for producing bio-image nanoparticles having excellent dispersibility in aqueous solution and preferential targeting by an introduction of an irregular surface structure in high yield, and according to the method of the present invention, hydrophobic nanoparticles. After partially modifying the surface of the particles with hydrophilicity, the functional groups are introduced into the hydrophilic group to produce functional nanoparticles having an irregular surface structure and including hydrophilic portions but exhibiting hydrophobicity as a whole, and then remaining hydrophobic portions of the surface of the nanoparticles. Is converted to hydrophilic to eliminate the agglomeration of hydrophilic nanoparticles and to ensure the independence and individuality of the particles throughout the entire reaction process to obtain bio-image nanoparticles having both aqueous phase dispersion, biocompatibility and target orientation. It can be prepared in yield.

Bio-imaging, Nanoparticles, Quantum Dots, Surface Modification, Biocompatibility, Target Orientation

Description

METHODS FOR THE PRODUCTION OF BIO-IMAGING NANOPARTICLES WITH HIGH YIELD BY EARLY INTRODUCTION OF IRREGULAR STRUCTURE}

The present invention relates to a method for producing bio-image nanoparticles having high dispersibility, biocompatibility and target orientation in aqueous solution by preferential introduction of irregular surface structures.

Recently, as a method of synthesizing hydrophobic inorganic nanoparticles having a uniform particle size by an organic method on an organic solution including a surfactant has been established, efforts have been actively made for practical use thereof. Particularly, in the case of nanoparticles synthesized in an aqueous solution, the particle size is much more uneven than that of the nanoparticles synthesized in an organic solution, and water is the cheapest, environmentally friendly, and most useful solvent present on the earth. Modification of the surface of the synthesized hydrophobic nanoparticles with uniform particle size into nanoparticles that can be stably dispersed in an aqueous solution is of great importance and is a field of interest for researchers. Among them, water-soluble quantum dots or water-soluble nanoparticles in the form of a single quantum dot or a single inorganic nanoparticle and a combination of organic materials having functional groups required on the surface thereof are not only bio-imaging materials but also biosensors and storage media. It is very useful as a base material for making a specific type of nanostructure, and thus research is being focused on surface modification.

The most widely used method for surface modification is the excessive reaction of an organic ligand in which thiol (SH) and a hydrophilic group are linked through a hydrocarbon chain to a hydrophobic nanoparticle, thereby all of the surfactant ligands on the surface of the nanoparticle are metal-thiolate (MS). After hydrophilic nanoparticles are prepared by substituting bonds and directing the hydrophilic groups of the organic ligand to the outside, a covalent bond between the hydrophilic groups of the nanoparticles and functional molecules such as a target-oriented biomolecule is formed to form a bio-containing nanoparticle containing only inorganic nanoparticles. To prepare a nanoparticle for the image (see Fig. 1 ). These covalent bonds mainly consist of amide bonds or ester bonds between the hydrophilic end groups of the nanoparticles and the new functional molecules. Polar organic ligands in which one of an amine (NH ₂ ) group, a carboxy (COOH) group, a thiol group and a hydroxy (OH) group, and a thiol group are connected through a hydrocarbon chain as a hydrophilic end group are mainly studied. Metal-rich quantum dots (e.g. CdSe, ZnS, or core / shell structured CdSe / CdS, CdSe / ZnS, etc.), precious metal nanoparticles (e.g. Au, Ag) or iron oxide magnetic nanoparticles It is known. However, among these, hydroxy end groups or amine end groups are easily aggregated and precipitated even in neutral or near neutral solutions, and thus no further studies have been conducted. On the other hand, since the carboxyl group is present in a large amount in an ionized state in neutral solution, it is excellent in dispersibility and solution stability and is widely used as a hydrophilic end group for connecting functional molecules to nanoparticles by amide bonds.

However, the method must go through the step of activating the carboxyl group on the surface of the nanoparticles in weakly acidic aqueous solution, in which a large number of nanoparticles aggregate and precipitate (WC Chan and S Nie, Science 281: 2016, 1998; Wen Jiang, et al ., Chem . Mater 18:. 872, 2006 ), and especially more severe the symptoms that occur when the magnetic nanoparticles is mungchigo precipitation. The agglomerated and precipitated nanoparticles are not only difficult to proceed with the next step of the functional molecules, but even if the functional molecules are in progress, the product is mainly produced in which the functional molecules are bound only to the surfaces of the agglomerated nanoparticles ( FIG. 1). See the figure on the right obtained from step B). The nanoparticles thus prepared are too large to move along the blood vessel and have significantly reduced dispersibility, and in the case of quantum dots, fluorescence is extremely reduced by self quenching. Eventually, the problem is that a large amount of nanoparticles is lost because these precipitates are separated and removed and only some well-dispersed solution layers are used.

In order to solve such agglomeration and precipitation phenomenon, recently, a hydrophobic inorganic nanoparticle is directly reacted with a polyethylene glycol [poly (ethylene glycol), PEG] polymer having a thiol end group and an organic-inorganic composite nanoparticle having an MS bond. It has been reported how to prepare (US Pat. No. 7,041,371). However, this method has a problem that the yield of the reaction is still low because the hydrophilic thiol group connected to the long chain must penetrate the surface of the nanoparticles surrounded by the surfactant.

Accordingly, the present inventors have diligently researched to solve the problems of the prior art in which hydrophilic nanoparticles are aggregated and precipitated in an aqueous solution. As a result, the hydrogen bonding force caused by the excess hydrophilic group on the surface of the hydrophilic nanoparticles having a regular structure is caused by the aggregation and precipitation. And the method of structurally obstructing the hydrogen bonding force between particles by securing the independence and individuality of the nanoparticles throughout the reaction process, without causing agglomeration or sedimentation. The present invention was completed by confirming that only nanoparticles containing only single particles and exhibiting excellent physical properties in particle uniformity, solution phase dispersibility and stability, biocompatibility, and target orientation can be prepared.

Accordingly, an object of the present invention is to produce nanoparticles for bio-imaging, by introducing an irregular structure on the surface of the nanoparticles by partial surface modification to structurally hinder the aggregation and sedimentation caused by hydrogen bonding forces between the particles. It is to provide a method for producing a bio-image nanoparticles having excellent dispersibility and stability in a non-polar organic solvent or an aqueous solution in accordance with the solvent in a high yield without loss of.

Another object of the present invention is to provide a nano-particle for bio-image having excellent dispersibility and stability produced by the above method.

In order to achieve the above object, the present invention

1) Substituting some of the surfactant by adding 1 to 30 equivalents of organic ligands in which the thiol group and the hydrophilic group are linked through a hydrocarbon chain selected from 8 to 20 carbon atoms to the hydrophobic nanoparticles having a surfactant or core / shell structure. And forming a metal-thiolate (MS) bond on the surface of the nanoparticles to produce hydrophobic nanoparticles in which only a portion of the surface is hydrophilically modified and maintains individual dispersibility in a nonpolar organic solvent;

2) introducing functional and irregular surface structures on the surface of the nanoparticles while maintaining individual dispersibility by binding functional molecules to the hydrophilic groups introduced to the surface of the nanoparticles prepared in step 1); And

3) converting the remaining surfactant remaining on the surface of the nanoparticles prepared in step 2) with an organic ligand connected with at least two hydrophilic groups through a hydrocarbon chain selected from 1 to 7 carbon atoms to convert them into hydrophilic nanoparticles It includes, it provides a method for producing a nano-particles for bio-images.

In addition, the present invention provides a metal-thiolate-bonded thiol group and a hydrophilic group selected from the group having 8 to 20 carbon atoms to the surface portion of the surfactant-protected core and core / shell structured hydrophobic inorganic nanoparticles prepared by the above method. Portions, including organic ligands linked by hydrocarbons, are hydrophilic but provide hydrophobic overall and nanoparticles for bio-images that maintain individual dispersibility in nonpolar organic solvents.

In addition, the present invention has a functional and irregular surface structure by the functional molecules are bonded to a hydrophilic group introduced to the surface of the nanoparticles prepared by the above method, the functional molecules are bonded to the hydrophilic part, but as a whole hydrophobic It provides a nano-particles for bio-image showing.

Finally, the present invention is hydrophilic by the remaining surfactant remaining on the surface of the nanoparticles prepared by the above method is substituted with an organic ligand connected to at least two hydrophilic groups through a hydrocarbon chain selected from 1 to 7 carbon atoms Provided are nanoparticles for bio-imaging that are converted and maintain individual dispersibility in aqueous solution.

The manufacturing method according to the present invention has a property of spherical shape of 20 nm or less, particle uniformity, chemical stability, aqueous phase dispersibility, biocompatibility, target orientation, etc., and has paramagnetic or high efficiency luminescence and light stability. The particles can be produced in 100% yield, and the nanoparticles produced by this method can be used as a bio-imaging material and can be used as a medical basic material that can be used to diagnose and treat diseases with high sensitivity. Can also be utilized. In addition, the manufacturing method of the present invention can be combined with a functional material through a hydrophilic group in the organic material, so long as the metal component on the surface of the nanoparticle forms a covalent bond with an organic material having a thiol regardless of the components of the internal inorganic nanoparticles. Various applications may be made in the preparation of organic-inorganic composite nanoparticles including various kinds of single nanoparticles.

Conventional surface modification method for producing nanoparticles for bio-imaging, hydrophilic nanoparticles made by binding a hydrophilic organic ligand to the surface of the hydrophilic nanoparticles in the aqueous solution or during the binding reaction with the biomolecules, the rules of surface hydrophilic functional groups The yield was extremely low due to the aggregation and precipitation caused by the hydrogen bonds induced by the structure.

Accordingly, the present inventors found that the particles of uniform and hydrophilic nanoparticles agglomerate and precipitate in an aqueous solution due to hydrogen bonding between particles caused by a regular structure of hydrophilic organic matter, and by stepwise partial surface modification, biocompatible molecules and targets are identified. Irregular structures of functional organics, such as directional molecules, combinations or mixtures thereof, are first introduced to produce nanoparticles that are hydrophobic, biocompatible, target-oriented, or both, and then convert the nanoparticles to hydrophilic. A method for producing nanoparticles for bio-images dispersed in an aqueous solution with high yield was developed by interfering hydrogen bonds between particles due to the steric hindrance of functional organics.

Specifically, the manufacturing method of the nano-particles for bio-image according to the present invention

1) Substituting some of the surfactant by adding 1 to 30 equivalents of organic ligands in which the thiol group and the hydrophilic group are linked through a hydrocarbon chain selected from 8 to 20 carbon atoms to the hydrophobic nanoparticles having a surfactant or core / shell structure. And forming a metal-thiolate (MS) bond on the surface of the nanoparticles to produce hydrophobic nanoparticles in which only a portion of the surface is hydrophilically modified and maintains individual dispersibility in a nonpolar organic solvent;

2) introducing functional and irregular surface structures on the surface of the nanoparticles while maintaining individual dispersibility by binding functional molecules to the hydrophilic groups introduced to the surface of the nanoparticles prepared in step 1); And

3) converting the remaining surfactant remaining on the surface of the nanoparticles prepared in step 2) with an organic ligand connected with at least two hydrophilic groups through a hydrocarbon chain selected from 1 to 7 carbon atoms to convert them into hydrophilic nanoparticles It includes.

As mentioned in the prior art, when hydrophobic nanoparticles with uniform particle sizes are synthesized in an organic solution containing a surfactant, and an MS bond is formed between the organic ligand and the nanoparticles in which the thiol group and the hydrophilic group are connected through a short hydrocarbon chain, Hydrophilic functional groups can be exposed to the outside to secure the hydrophilicity of the powder. However, the hydrophilic nanoparticles exposed to the hydrophilic group are exposed to agglomerate so much that amide or ester bond formation with other biopolymers cannot be successfully performed. For this reason, there have not been reported examples of producing nanoparticles for bio-images containing single nanoparticles at the center in high yield.

In the present invention, as shown in FIG. 1 , various hydrophilic nanoparticles including hydrophilic nanoparticles exposed with an amine group or a carboxy group are self-assembled on the surface of the particles regularly, and thus, interparticle hydrogen by an excessive amount of hydrophilic functional groups. Due to the binding force, it was found to be agglomerated and settled. In order to suppress such a phenomenon, the present invention excludes a phenomenon in which particles are aggregated and precipitated by fundamentally hindering hydrogen bonds between particles by making irregular structures on the surface of the nanoparticles through a partial partial surface modification as shown in FIG. 2 . The first conditions that can be distributed individually were established. Under these conditions, the hydrophobic nanoparticles first replace only some of the surfactants with organic ligands in which a thiol group and a hydrophilic group are connected through a hydrocarbon chain selected from 8 to 20 carbon atoms to form an MS bond, and the hydrophilic group has biocompatibility and target orientation. By combining functional molecules, they have a functional and irregular structure, and hydrophobic functional nanoparticles in this state are themselves in It can be used for special purpose bio-imaging such as in vitro cell experiments. Subsequently, the remaining unsubstituted surfactant is converted into hydrophilic nanoparticles by replacing at least two hydrophilic groups with organic ligands connected through a hydrocarbon chain selected from 1 to 7 carbon atoms, whereby the hydrogen bonds between the particles are structurally disturbed. The nanoparticles for bio-images with improved dispersibility and stability in aqueous phase including single nanoparticles can be prepared in high yield.

Referring to Figure 2 , it will be described step by step the manufacturing method of the nano-particles for bio-image according to the present invention.

Step 1) is a partial surface modification step of the hydrophobic nanoparticles (see C of FIG. 2 ), which includes inorganic nanoparticles of core 10 or core / shell 12 structure protected by surfactant 14. A small amount, preferably 1 to 30 equivalents, of the organic ligand 20 in which a thiol group and a hydrophilic group forming a chemical bond with a metal element on the surface of the nanoparticles in the organic solution are connected through a hydrocarbon chain selected from 8 to 20 carbon atoms. Add and react with vigorous stirring. Through this reaction, some surfactants on the surface of the nanoparticles are substituted with the organic ligands, and the substituted organic ligands form MS covalent bonds with metal elements on the surface of the nanoparticles.

In step 1), the inorganic nanoparticles are core or core / shell structures, and the surface of the nanoparticles is more stoichiometrically richer than metals, and the metals are chemically formed by MS covalent bonds with thiol groups, which are members of organic ligands. Can be combined. The inorganic nanoparticles are semiconductor nanoparticles, precious metal nanoparticles or iron oxide magnetic nanoparticles composed of one element of zinc, cadmium, and lead, which are group II elements on the periodic table, and one element of sulfur, selenium, and tellurium, which are group VI elements on the periodic table. Is preferably. More preferably, examples of the inorganic nanoparticles include CdSe, ZnS, CdSe / CdS, CdSe / ZnS, PbS, Au, Ag, Fe ₂ O ₃ , Fe ₃ O ₄ , and the like. Nanoparticles composed of a material that can be formed can be used without limitation in the present invention.

The organic ligand binding to the inorganic nanoparticles may include at least one thiol group and at least one hydrophilic group in a molecule to form at least one MS bond with the inorganic nanoparticles, and may be formed of another biopolymer 18 such as a bioffin. At least one binding site can be provided for binding to a chemical molecule, a target oriented molecule, a conjugate thereof (bioaffinity-target oriented molecule) or a mixture.

In addition, the length of the long hydrocarbon chain in the organic ligand is preferably 8 to 20 to maintain the hydrophobicity of the nanoparticles. If the length of the hydrocarbon chain is less than eight nanoparticles may lose hydrophobicity, the chain length is too short may cause the hydrophilic group is hidden in the surfactant it is difficult to form a bond with the functional molecule. On the other hand, organic ligands are not yet developed when the length of the hydrocarbon chain exceeds 20, but if longer organic ligands are developed, they can be used. At this time, the amount of the organic ligand added is preferably in the range of 1 to 30 equivalents. If less than 1 equivalent is added, the original nanoparticles in which the organic ligand is not substituted at all are present. When added in excess of 30 equivalents, the hydrophilic group is excessively substituted on the surface of the nanoparticles, resulting in a strong hydrogen bonding force may cause the nanoparticles to aggregate and precipitate.

The number of organic ligands that can be chemically bonded by M-S covalent bonding between a metal element and a thiol group on the surface of the inorganic nanoparticle may vary depending on the size of the particles, and may be 1 to 150. However, when more than 30 M-S bonds were made and the biocompatible polymers were bound thereto, the particles aggregated to form aggregates and were not dispersed in any solvent. Therefore, the number of organic ligands that can be chemically bonded by M-S covalent bonds between metal elements and thiol groups on the surface of the inorganic nanoparticles is preferably 1 to 30. Examples of organic ligands that can be used to form irregular structures on the surface of nanoparticles include mercaptoundecanoic acid, mercaptododecanoic acid, and mercaptohexadecanoic acid. .

Partially surface-modified inorganic nanoparticles at this stage are hydrophobic, although the hydrophilic group is exposed to the outside, the length of the hydrocarbon chain to which the hydrophilic group is linked is substantially non-hydrophilic, and the nanoparticles are still hydrophobic because the number is much less than hydrophobic surfactants. And disperse well in nonpolar solvents.

Step 2) binds a functional molecule such as a biocompatible molecule, a target-oriented molecule, a conjugate or a mixture thereof to the hydrophilic group of the nanoparticle prepared in step 1), and has a nanostructure having an irregular structure by the functional molecule introduced to the surface. To prepare a step. The nanoparticles produced at this stage are still hydrophobic or amphoteric, well dispersed in nonpolar solvents and in It can be used for special purpose bio-imaging such as in vitro cell experiments.

Step 3) adds an organic ligand (16) in which at least two hydrophilic groups are linked through a hydrocarbon chain selected from 1 to 7 carbon atoms to the nanoparticles prepared in step 2) to replace the surfactant remaining on the surface of the nanoparticles. By converting to hydrophilic nanoparticles. Examples of suitable organic ligands suitable for this step include mercaptohexanoic acid, mercaptoacetic acid, mercaptopropionic acid, dimercaptosuccinic acid, and 2-mercaptoethanol (2). mercaptoethanol, 2-aminoethanethiol, lysine, arginine, aminovaleric acid, and the like.

The irregular structure introduced on the surface of the nanoparticles through steps 1) and 2) inherently inhibits hydrogen bonding between the hydrophilic nanoparticles that may occur in step 3), thereby maintaining individual dispersibility, and introduced in step 3). The short hydrocarbon chain length selected from 1 to 7 carbon atoms of the organic ligands increases the polarity of the organic material and serves to improve dispersibility in water.

In the present invention, the functional molecule is a synthetic polymer or a bioconstituent material, and has excellent biocompatibility, and has no side effects, and means a material having various physical properties such as solution, gel, and membrane depending on biodegradability and conditions, and at least one terminal includes an amine group and an aldehyde. A material having one of a group and a carboxyl group is to form an amide or ester bond with the hydrophilic group of the nanoparticle. Examples of such functional molecules include biocompatible molecules, target oriented molecules, conjugates thereof (biocompatible-target oriented molecules) or mixtures thereof.

The biocompatible molecule has a group selected from an amine group, an aldehyde group and a carboxy group at both chain ends, or has one of an amine group, an aldehyde group and a carboxy group at one chain end, and has 1 to 7 carbon atoms at the other chain end. It is preferable to have an alkoxy group or a hydroxyl group selected from dogs. Such biocompatible molecules include polyethylene glycol (PEG), dextran, poly (L-lactide) (PLLA), poly (DL-lactide) (PDLLA), poly-DL-lactide / glycolide aerial Copolymer (PLGA), chitosan, alginic acid, hyaluronic acid, collagen, heparin, poly (ε-caprolactone) and the like can be used.

In addition, the target-oriented molecule has a amine group, a carboxyl group, a hydroxy group and a thiol group as a substance specifically recognized in vivo, and thus the amide bond, ester bond and thio group with the organic ligand or the biocompatible molecule. It is preferred that they can be linked by one of the ester bonds. Folic acid or methotrexate (MTX), which selectively reacts with folate receptor proteins with these target-directed molecules, peptides that are selective for specific cells, such as RGD peptides or tat peptides (tat peptides), or antibodies that selectively react to specific antigens, such as biotin against streptavidin, PSA antibodies against PSA, and the like.

In addition, a biocompatible-targeted molecule or a mixture thereof prepared by combining the biocompatible molecule and the target-oriented molecule by an amide bond or an ester bond may be used.

Functional molecules such as biocompatible molecules, target oriented molecules, biocompatible-target oriented molecules or mixtures thereof have at least one of amine groups, aldehyde groups, carboxy groups and hydroxy groups at one end of the molecule. In step 2), an amide bond or an ester bond with the carboxy group, the hydroxyl group or the amine group of the individually dispersed nanoparticles may be formed to form nanoparticles having an irregular structure of organic matter on the surface.

As shown in step B of Figure 1 , the conventional method for producing nanoparticles is a form in which the functional molecules are surrounded only on the surface of the population because the hydrophilic nanoparticles perform a biopolymer and amide or ester coupling reaction in a clustered state As the nanoparticles are formed and the nanoparticles present therein cannot be contacted with the functional molecules, they cannot form a bond. This has resulted in lower yields and lower imaging capabilities of the final bio-imaging nanoparticles.

However, in the method for preparing nanoparticles according to the present invention, as shown in step D of FIG . 2 , a portion of the surfactant on the surface of the nanoparticles is selected from 8 to 20 carbon atoms by staged partial surface modification. Substituted with organic ligands connected through the functional molecules bonded to the organic ligands to prepare a hydrophobic functional nanoparticles having an irregular structure first. Thereafter, the remaining surfactant on the surface of the nanoparticles is replaced with an organic ligand connected to at least two hydrophilic groups through a hydrocarbon chain selected from 1 to 7 carbon atoms to convert the hydrophobicity of the nanoparticles to hydrophilicity. According to the manufacturing method of the present invention, since the aggregation and precipitation due to the hydrogen bonding between particles are not caused by the irregular structure introduced on the surface of the nanoparticles, they include a single nanoparticle in the center without loss of hydrophilic quantum dots. Organic-inorganic composite nanoparticles can be prepared in 100% yield as well as conserving quantum efficiency.

According to a preferred embodiment of the present invention, a core or core / shell nanoparticle having a spherical shape and a fluorescent property of 20 nm or less, or iron oxide magnetic nanoparticles are synthesized in an organic solution to ensure uniformity of the particles. By combining the long-chain organic ligand and the functional molecules in turn by surface modification, nanoparticles for irregular hydrophobic functional bio-imaging are provided. In addition, nano-particles for bio-images that maintain individual dispersibility in aqueous solution by converting hydrophobicity of the nanoparticles into hydrophilic state in the state that steric hindrance due to irregular structure on the surface of the nanoparticles interferes with hydrogen bonding between particles to be operated later. In 100% yield.

In addition, the manufacturing method of the present invention, regardless of the constituents of the inorganic nanoparticles inside the nanoparticles surface is bound to the functional molecule through the hydrophilic group in the organic ligand as long as the metal component of the nanoparticles to form a covalent bond with the organic ligand having a thiol group. Therefore, it can be usefully applied to the production of organic-inorganic composite nanoparticles including not only quantum dots and magnetic nanoparticles but also various types of single nanoparticles.

In addition, in the preparation method of the present invention, even though the covalent bond between the inorganic nanoparticle and the organic ligand is not an MS bond, if the amine group, the carboxyl group, or the hydroxyl group is exposed at the end of the organic ligand substituted by the step partial surface modification, the functional molecule is a functional molecule. It can be converted into hydrophilic form after forming a structure that steric hindrance interferes with hydrogen bonding between particles by forming a bond with, so that it can be applied to the production of various kinds of discrete bio-image nanoparticles before and after hydrophilic conversion. Can be.

The present invention also provides a nano-particle for bio-imaging comprising a single inorganic nanoparticles in the center produced by the method.

The nanoparticles for bio-imaging according to the present invention include an organic ligand having a long hydrocarbon chain, which is MS-bonded to a portion of the surface of an inorganic nanoparticle having a core and a core / shell structure, and the organic ligand-bonded portion is hydrophilic. And most of the others include hydrophobic nanoparticles (produced by step 1) in the method of the present invention, wherein the hydrophilic groups of the nanoparticles are connected to functional molecules by amide or ester bonds and have irregular surface structures. Hydrophobic nanoparticles (produced by step 2) in the preparation method of the present invention, and the remaining surfactant remaining on the surface of the nanoparticles is replaced with a polar organic ligand of a short hydrocarbon chain having at least two hydrophilic groups to be hydrophilic. Hydrogen bonds between particles are prevented by the steric hindrance of irregular surface structures. It comprises ah through the full cycle of the reaction hydrophilic nanoparticles maintain the individual dispersion produced by the (step 3 of the method of the present invention)).

The hydrophilic bio-imaging nanoparticles according to the present invention have a large number of hydrophilic organic ligands capable of hydrogen bonding on one nanoparticle surface, but the functionalities bound to organic hydrocarbons of long hydrocarbon chains providing irregular structures on some surfaces. Since the molecules are bonded, they give a steric hindrance to hydrogen bonding between particles, and they have excellent particle uniformity, chemical stability, aqueous phase dispersion, biocompatibility, and target orientation without loss of nanoparticles due to aggregation and precipitation. Luminous properties and light stability can be secured.

Therefore, the nanoparticles according to the present invention may not only be very useful as a bio-imaging material but may also be used as a medical base material capable of carrying out the diagnosis and treatment of diseases with high sensitivity.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention more specifically, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention.

The hydrophobic nanoparticles CdSe / CdS-ODA of the core / shell structure used as starting materials of the quantum dots in the following examples are coordinated with octadecylamine (ODA) surfactants on the entire surface, and described in JJ Li, et al., Journal of the American Chemical Society 125: 12567-12575, 2003; WW Yu, et al., Chemistry A total of 2.5 layers of Cd, S, Cd, S, and Cd were sequentially grown on the surface of the CdSe core nanoparticles, respectively, in turn, by referring to the method described in of Materials 15: 2854-2860, 2003).

In addition, the hydrophobic iron oxide nanoparticles Fe ₂ O ₃ -OA used as starting materials of the magnetic nanoparticles in the following examples are coordinated with an oleic acid (OA) surfactant on the entire surface of the nanoparticles (K. Woo, et al., Chemistry of Materials 16: 2814-2818, 2004; K. Woo, et al., IEEE Transactions on magnetic particles 41, 4137-4139) and synthesized Fe ₂ O ₃ or Fe ₃ O ₄ nanoparticles in an inert atmosphere by adding 10 mol% precursor Fe (CO) ₅ and refluxing the amount of iron on the surface The theoretically rich magnetic nanoparticles were prepared. However, it will be apparent to those skilled in the art that any method can be applied to the same as in Examples 1 to 11 as long as the stoichiometrically rich hydrophobic nanoparticles of the metal component on the surface.

In addition, in the following examples, (-DA) _ex or (-OA) _ex is an abbreviation indicating an excess of decylamine (DA) or oleic acid (OA) on the surface of nanoparticles, in which some ligands are substituted with ligands having a thiol group. Even if the reduction effect is minimal and there is still an excess of DA or OA, they will be referred to as (-DA) _ex or (-OA) _ex without distinguishing them.

Example  1: surface Ligand  Semiconductor Nanoparticles by Exchange CdSe Of CdS - DA Manufacture

Take 5 ml of hydrophobic CdSe / CdS-ODA quantum dot solution (8 × 10 ^-5 M), connect to vacuum, remove solvent, disperse in 20 ml of chloroform, add 1000 equivalents of decylamine (DA) It stirred for 2 days in atmosphere. The precipitate formed by adding acetone to this solution was centrifuged and dispersed in chloroform to prepare 20 ml of a CdSe / CdS-DA solution (2 × 10 ⁻⁵ M). We analyzed the CdSe / CdS-DA sample the infrared spectrum and transmission electron microscopy (transmission electron microscope, TEM) images of, and the results are shown in the (b) of Fig. 4 (b) of Figure 3, respectively. In the TEM image, the interval between the quantum dots was shorter than that of CdSe / CdS-ODA, indicating that ligand exchange occurred.

Example  2: partial surface Modified  Semiconductor Nanoparticles CdSe Of CdS (- DA ) _ex (- MUA ) ₅ Manufacture

17 ml of the CdSe / CdS-DA solution prepared in Example 1 was taken, 5 equivalents of mercaptoundecanoic acid (MUA) was added, and the mixture was stirred for 19 hours under a dark inert atmosphere. After the solution was concentrated, acetone was added, and the resulting precipitate was centrifuged and dispersed in chloroform to prepare 17 ml of a CdSe / CdS (-DA) _ex (-MUA) ₅ solution (2 × 10 ⁻⁵ M). We analyzed the infrared spectra and TEM images of the samples, and the results are shown in the (c) of Figure 4, respectively (c) of Fig. It was confirmed in the TEM image that the self-assembly structure no longer appeared as the rule structure was broken as in step C of FIG. 2 due to partial substitution of the MUA.

Example  3: Target-Oriented Hydrophobic Semiconductor Nanoparticles CdSe Of CdS (- DA ) _ex (- MUA - en - FA ) ₅ Manufacture

2 ml of a solution of CdSe / CdS (-DA) _ex (-MUA) ₅ prepared in Example 2 was taken and diluted to 10 ml by addition of chloroform. After adding 5 equivalents of dicyclohexylcarbodiimide (DCC) and stirring for 3 hours in a dark inert atmosphere, 50 equivalents of en-FA prepared as follows was added and further stirred for 2 hours. After concentrating the solution, the resultant precipitate was added by the addition of acetone and centrifuged and dispersed in chloroform to dissolve 10 ml of CdSe / CdS (-DA) _ex (-MUA-en-FA) ₅ solution (4 × 10 ^-6 M). Was prepared. en-FA was the analysis that the binding to the MUA in the infrared spectrum and the TEM image, The results are shown in (d) of the Figure 4 (d) of Figure 3, respectively.

Target oriented molecule folic acid ( folic acid , FA )and Of ethylenediamine  concrete en - FA Manufacture

441 mg (1 mmol) of folic acid was added to 10 ml of dry distilled toluene, stirred well for 1 hour, and then connected to a vacuum to remove toluene and water. This was dissolved in dimethylformamide (DMF), and the flask was placed in an iced water bath and stirred for 10 minutes. To this was added 226 mg (1.1 mmol) of DCC and further stirred for 18 hours. 2.5 ml of ethylenediamine (en) was dissolved in 20 ml of DMF and the solution was added and stirred for 3 hours. Water and acetonitrile were added to the solution to form a precipitate, which was recrystallized from the same solvent to obtain 224 mg of en-FA.

Example  4: Biocompatible Water Soluble Semiconductor Nanoparticles CdSe Of CdS (- MPA ) _ex (- MUA - aPEGa ) ₅ Manufacture

0.5 mL of the CdSe / CdS (-DA) _ex (-MUA) ₅ solution prepared in Example 2 was taken, and diluted with 2.5 mL by adding chloroform. To this was added 6 equivalents of DCC and stirred under dark inert atmosphere for 3 hours, followed by O, O'-bis (2-aminoethyl) polyethylene glycol (O, O'-Bis (2-aminoethyl) polyethylene glycol, 7 equivalents of aPEGa, molecular weight 1,000) was added and further stirred for 16 hours. The solution was concentrated and acetone was added to centrifuge the resulting precipitate and dispersed in chloroform to prepare 10 ml of a solution of CdSe / CdS (-DA) _ex (-MUA-aPEGa) ₅ (1 × 10 ^-6 M). It was.

Subsequently, 100 equivalents of methanol solution in which 0.05 M NaOH and 0.05 M mercaptopropionic acid (MPA) were dissolved and added to the CdSe / CdS (-DA) _ex (-MUA-aPEGa) ₅ solution was stirred. Distilled water was added thereto, the product was extracted, and the resultant precipitate was centrifuged by adding a methanol / ethyl acetate (1/4) mixed solvent. The separated precipitate was dispersed in PBS buffer at pH 7.4 to prepare 10 ml of 1 × 10 ⁻⁶ M CdSe / CdS (−MPA) _ex (-MUA-aPEGa) ₅ . aPEGa is coupled to the MUA and DA were analyzed via a substitution that infrared spectra and TEM images to MPA, the results are shown in (e) of the Fig. 4 (e) of Figure 3, respectively.

Example  5: Target Oriented-biocompatible Water-Soluble Semiconductor Nanoparticles CdSe Of CdS (- MPA ) _ex (- MUA - aPEGa - FA ) ₅ Manufacturing and bio- Imaging

2 ml of a solution of CdSe / CdS (-DA) _ex (-MUA) ₅ prepared in Example 2 was taken and diluted to 10 ml by addition of chloroform. After 6 equivalents of DCC was added thereto and stirred for 3 hours in a dark inert atmosphere, 15 equivalents of aPEGa-FA prepared as follows was added thereto, followed by further stirring for 16 hours. The solution was concentrated and acetone and methanol were added to the resulting precipitate. The precipitate was centrifuged and dispersed in chloroform to give a solution of CdSe / CdS (-DA) _ex (-MUA-aPEGa-FA) ₅ (1 × 10 ^-5 M). 4 ml were prepared.

Subsequently, 100 equivalents of a methanol solution in which 0.05 M NaOH and 0.05 M mercaptopropionic acid (MPA) were dissolved and stirred was added to the CdSe / CdS (-DA) _ex (-MUA-aPEGa-FA) ₅ solution. Distilled water was added thereto, the product was extracted, and the resultant precipitate was centrifuged by adding a methanol / ethyl acetate (1/4) mixed solvent. The separated precipitate was dispersed in PBS buffer at pH 7.4 to prepare ₅ 10 ml of 4 × 10 ⁻⁶ M CdSe / CdS (-MPA) _ex (-MUA-aPEGa-FA). is coupled to the MUA aPEGa-FA and DA were analyzed via substitution that infrared spectra and TEM images in MPA, given in (f) of Figure 4 and (f) of Figure 3 and the results, respectively.

In order to check whether FA-conjugated quantum dot CdSe / CdS (-MPA) _ex (-MUA-aPEGa-FA) ₅ shows selectivity to specific cells expressing folic acid receptors, KB cells with folic acid receptors ( Human fibrosarcoma cells (ATCC Manassas, VA) and HT1080 cells without folic acid container (Human epidermoid carcinoma cells (ATCC Manassas, VA)) were combined with CdSe / CdS with FA in the presence and absence of FA (9 × 10 ^-6 M) ( -MPA) _ex (-MUA-aPEGa-FA) ₅ quantum dots (2 × 10 ^-7 M) or CdSe / CdS (-MPA) _ex (-MUA-aPEGa) ₅ quantum dots (2 × 10 ^-7 without FA) Inoculated in PBS solution containing M) and incubated for 15 hours at 37 ° C. After the incubation was completed, the cells were washed three times with PBS solution and fixed on a slide glass. The results are shown in FIG. 5 .

As shown in FIG . 5 , when there is no free FA in the culture, the target-oriented quantum dot CdSe / CdS (-MPA) _ex (-MUA-aPEGa-FA) ₅ has no target-oriented quantum dot CdSe / CdS (-MPA) _ex ( -MUA-aPEGa) ₅ much larger amounts were selectively delivered into KB cells with FA receptors, whereas in the presence of excess FA in the culture, free FA binds most of the receptors between KB cells and HT1080 cells. It was confirmed that no selectivity appeared.

Biocompatible molecules aPEGa And target-oriented molecules FA By the combination of aPEGa - FA Manufacture

441 mg (1 mmol) of FA was added to 10 ml of dry distilled toluene and stirred well for 1 hour, followed by vacuum to remove toluene and water. This was dissolved in 14 ml of dimethylformamide and the flask was placed in an iced water bath and stirred for 10 minutes. 226 mg (1.1 mmol) of DCC was added thereto, followed by stirring for 18 hours, followed by 1 mmol of aPEGa, followed by further stirring for 3 hours. Diethyl ether was added to this solution to form a precipitate, which was recrystallized from the same solvent to obtain 179 mg of aPEGa-FA.

Example  6: semiconductor nanoparticle CdSe Of CdS (- DA ) _ex (- MUA - aPEGa ) _n Preparation of (n = 5, 10 or 30)

0.5 ml of the CdSe / CdS-DA solution prepared in Example 1 was taken, and three solutions diluted to 10 ml were prepared by adding chloroform. 5, 10 and 30 equivalents of mercaptoundecanoic acid (MUA) were added to the solution and stirred for 19 hours in a dark inert atmosphere. The solution was concentrated and methanol was added to the resulting precipitate, which was then centrifuged and dispersed in chloroform to give CdSe / CdS (-DA) _ex (-MUA) _n (n = 5, 10 or 30) solution (4 × 10 ^{−). 6} M) 2.5 ml each was prepared.

CdSe / CdS (-DA) _ex (-MUA) _n (n = 5, 10 or 30) above 5.5, 11 and 33 equivalents of DCC were added to the solution and stirred for 3 hours in a dark inert atmosphere, and then 6, 12 and 36 equivalents of aPEGa were added to each solution and further stirred for 16 hours. After concentration of this solution, the resultant precipitate was centrifuged by the addition of methanol and dispersed in chloroform to give a solution of CdSe / CdS (-DA) _ex (-MUA-aPEGa) _n (n = 5, 10 or 30) (1 ×). 10 ^-6 M) 10 ml each was prepared. In all three cases, it was confirmed by infrared spectra that aPEGa was bound to MUA, and the results are shown in FIG. 6 .

In addition, the effect of hydrogen bonding force between nanoparticles is small in the case of CdSe / CdS (-DA) _ex (-MUA-aPEG) ₅ through TEM image, while CdSe / CdS (-DA) _ex (-MUA-aPEG) _In the case of ₁₀ it was confirmed that in Figure 7 to act significantly to form agglomerated nanoparticles. In the above formula, when n is 5 and 10, it is well dispersed in chloroform, but in case of 30, hydrogen bonding force between nanoparticles is too large to form a coagulated solid. No image was obtained. From this, it was confirmed that as the number of hydrophilic groups having a regular structure on the surface of the nanoparticles increases, the hydrogen bonding force between particles may be uncontrollably large.

In Examples 1 to 6 above, it was confirmed that fluorescence was not detected at all in the discarded liquid by adding an insoluble solvent to the quantum dot solution after the reaction and then centrifuging it to confirm that the yield was 100%.

Example  7: partial surface Modified  Magnetic Nanoparticles SPION (- OA ) _ex (- MHA ) ₁₀ Manufacture

2 ml of a 8 nm hydrophobic SPION-OA magnetic nanoparticle solution (3 × 10 ⁻⁶ M) was taken, washed twice with ethanol, and the solution dispersed in 20 ml of toluene was heated to 100 ° C. 10 equivalents of mercaptohexadecanoic acid (MHA) was added thereto, refluxed for 1 hour and cooled. After 2 ml of this solution was concentrated, a small surface-modified magnetic nanoparticle SPION (-OA) _ex (-MHA) ₁₀ was separated using a magnet with a small amount of ethanol, and its infrared spectroscopy and TEM analyze the image shown in (b) of Fig. 9 and Fig. 8 (b), respectively. In the TEM image, the spacing between the quantum dots of SPION (-OA) _ex (-MHA) ₁₀ is similar to that of SPION-OA, where MHA, which has two carbon chains shorter than OA, is sporadic as the product obtained in step C of FIG. It was interpreted that the OA was partially substituted, and MHA was substituted in the infrared spectral spectrum, and it was confirmed that the OH peak near 3300 cm ⁻¹ was increased.

Example  8: Target-Oriented Biocompatible Hydrophobic Magnetic Nanoparticles SPION (- OA ) _ex (- MHA - aPEGa - MTX ) ₁₀ Manufacture

30 equivalents of DCC was added to 18 ml of SPION (-OA) _ex (-MHA) ₁₀ solution (3 × 10 ^-7 M) prepared in Example 7, followed by stirring for 3 hours, and then 30 equivalents of aPEGa-MTX prepared as follows. Was dissolved in a small amount of DMF and added and stirred for 16 h. After concentrating the solution and adding ethanol, SPION (-OA) _ex (-MHA-aPEGa-MTX) ₁₀ was separated using a magnet and dispersed in 9 ml of chloroform. Take a 1 ㎖ of the infrared spectrum was carried out with a TEM image analysis, and the results are shown in (c) of FIG. 9 (c) of Figure 8, respectively. In the TEM image, the magnetic nanoparticles were found to be scattered sporadically by breaking the hexagonal arrays by self-assembly, indicating that the irregular structure was introduced by aPEGa-MTX, and PEG and MTX in the infrared spectral spectrum near 1100 and 1630 cm ^-1 . It was confirmed that the characteristic peak of appeared.

Biocompatible molecules aPEGa And target-oriented molecules MTX By the combination of aPEGa - MTX Manufacture

454 mg (1 mmol) of MTX (methotrexate, USP Reference Standard) was dissolved in dry distilled 14 ml of dimethylformamide, and the flask was placed in an iced water bath and stirred for 10 minutes. 226 mg (1.1 mmol) of DCC was added thereto, followed by stirring for 18 hours, followed by 1 mmol of aPEGa, followed by further stirring for 3 hours. Diethyl ether was added to the solution to form a precipitate, which was recrystallized from the same solvent to obtain 138 mg of aPEGa-MTX.

Example  9: Target-Oriented Bio-Soluble Magnetic Nanoparticles SPION (- MPA ) _ex (- MUA - aPEGa - MTX ) ₁₀ Manufacture

150 equivalents of methanol solution dissolved in 0.05 M MPA and 0.05 M NaOH was added to 8 ml of SPION (-OA) _ex (-MHA-aPEGa-MTX) ₁₀ solution (6 × 10 ^-7 M) prepared in Example 8. Stir for 2 hours. Methanol was added thereto, and the solid SPION (-MPA) _ex (-MUA-aPEGa-MTX) ₁₀ obtained by centrifugation was dispersed in 16 ml of PBS buffer at pH 7.4. Take a 1 ㎖ of the analyzed infrared spectra and TEM images, and the results are shown in (d) (d) of Figure 8 and Figure 9, respectively. It was confirmed that the intensity of the CH peak was significantly reduced because the length of the carbon chain decreased as OA was substituted with MPA in the infrared spectroscopic spectrum. In addition, in the TEM image, it was confirmed that the distance between particles was secured even though there was an overlap effect due to the surface tension due to the use of an aqueous solution when preparing the specimen.

Example  10: Target-Oriented Biocompatible Water-Soluble Magnetic Nanoparticles SPION (- Lys ) _ex (- MUA - aPEGa - MTX ) ₁₀ Manufacture

1,000 equivalents of tetraoctylammonium bromide (TOAB) was added to 6 ml of ₁₀ solutions (6 × 10 ^-7 M) of SPION (-OA) _ex (-MHA-aPEGa-MTX) prepared in Examples 7 and 8. Stir for hours. 6 ml of 0.1 M aqueous solution of Lysine (Lysine, Lys) was added thereto, followed by stirring for 19 hours. Methanol was added to the solution, and SPION (-Lys) _ex (-MUA-aPEGa-MTX) ₁₀ nanoparticles were separated using a magnet, washed with 5 ml of toluene and 5 ml of ethanol, and a pH 7.4 PBS buffer solution. Dispersed in. Take a 1 ㎖ of the analyzed infrared spectra and TEM images, and the results are shown in the (e) (e) of Figure 8 and Figure 9, respectively. It was confirmed that the intensity of the CH peak was significantly reduced because the length of the carbon chain was reduced as OA was substituted with lysine in the infrared spectroscopic spectrum. In addition, in the TEM image, it was confirmed that the distance between particles was secured even though there was an overlap effect due to the surface tension due to the use of an aqueous solution when preparing the specimen.

Example  11: Target-Oriented Biocompatible Hydrophobic Magnetic Nanoparticles SPION (- OA ) _ex (- MHA - en - FA ) ₅ Manufacturing and bio- Imaging

2 ml of 11 nm hydrophobic SPION-OA magnetic nanoparticle solution (3 × 10 ⁻⁶ M) was taken, washed twice with ethanol, and the solution dispersed in 20 ml of toluene was heated to 100 ° C. 5 equivalents of mercaptohexadecanoic acid (MHA) was added thereto, refluxed for 1 hour and cooled. After 15 equivalents of DCC was added to the solution and stirred under a dark inert atmosphere for 3 hours, 15 equivalents of en-FA prepared in Example 3 was dissolved in a small amount of DMF and stirred for 16 hours. After concentrating the solution and adding ethanol, SPION (-OA) _ex (-MHA-en-FA) ₅ was separated using a magnet and dispersed in 10 ml of chloroform. The infrared spectral spectrum of this nanoparticle is shown in FIG. 8 (f), and the characteristic peak of FA was confirmed in the vicinity of 1630 cm ⁻¹ .

The prepared solution of SPION (-OA) _ex (-MHA-en-FA) ₅ was diluted to 1 × 10 ^-7 M, and 1 ml was taken from the solution and spread evenly on a culture dish, followed by natural drying. The next day, the plate was sterilized with 70% ethanol and UV light, and 2 × 10 ⁵ KB cells (ATCC Manassas, VA), which are human epithelial cancer cells, were inoculated thereon and incubated in a 37 ° C. incubator for 16 hours. The cultured cells were scraped and further incubated for 4 hours in a culture dish without any treatment. In order to compare the targeting behavior of the nanoparticles, the same experiment as above was performed except that 10 μg / ml FA was added to the culture solution. Magnetic resonance imaging (MR) images of each of the cells treated as described above were analyzed, and the results are shown in FIG. 10 . Quantitative analysis of the MR images revealed that coexistent excess free FA occupied the receptors on the cell surface, indicating that the target-oriented magnetic nanoparticle SPION (-OA) _ex (-MHA-en-FA) ₅ It was confirmed that the decrease of about 10%, from this it can be seen that the target orientation by FA of hydrophobic magnetic nanoparticles is a significant level.

In the above, for example, the nanoparticles for bio-images having irregular surface structures by partial surface modification and having some or all of hydrophilicity, biocompatibility, and target orientation have been described in detail. However, the foregoing embodiments are only presented by way of example to illustrate the present invention in detail, and the present invention is not limited to these examples unless explicitly stated in the following claims.

Figure 1 shows the manufacturing process and nanoparticle structure of each step of the nano-particles for bio-image according to the prior art,

Figure 2 shows the manufacturing process and nanoparticle structure of each step of the bio-image nanoparticles according to the present invention,

Figure 3 shows an infrared spectroscopy spectrum of the quantum dot starting material and the quantum dots prepared in Examples 1 to 5,

a) CdSe / CdS-ODA quantum dots used as starting material

b) CdSe / CdS-DA quantum dots prepared in Example 1

c) CdSe / CdS (-DA) _ex (-MUA) ₅ quantum dots prepared in Example 2

d) CdSe / CdS (-DA) _ex (-MUA-en-FA) ₅ quantum dots prepared in Example 3

e) CdSe / CdS (-MPA) _ex (-MUA-aPEGa) ₅ quantum dots prepared in Example 4

f) CdSe / CdS (-MPA) _ex (-MUA-aPEGa-FA) ₅ quantum dots prepared in Example 5

Figure 4 shows a transmission electron microscope (TEM) image of the quantum dot starting material and the quantum dots prepared in Examples 1 to 5,

a) CdSe / CdS-ODA quantum dots used as starting material

b) CdSe / CdS-DA quantum dots prepared in Example 1

c) CdSe / CdS (-DA) _ex (-MUA) ₅ quantum dots prepared in Example 2

d) CdSe / CdS (-DA) _ex (-MUA-en-FA) ₅ quantum dots prepared in Example 3

e) CdSe / CdS (-MPA) _ex (-MUA-aPEGa) ₅ quantum dots prepared in Example 4

f) CdSe / CdS (-MPA) _ex (-MUA-aPEGa-FA) ₅ quantum dots prepared in Example 5

Figure 5 shows the fluorescence microscope image of each cell after delivery to HT1080 cells and KB cells using the quantum dots prepared in Examples 4 and 5 for control and target orientation,

QD: CdSe / CdS (-MPA) _ex (-MUA-aPEGa) ₅ quantum dots used as controls

QD-FA: CdSe / CdS (-MPA) _ex (-MUA-aPEGa-FA) ₅ quantum dots used for targeting

+ FA: the presence of excess free FA

FA: free FA does not exist

6 shows an infrared spectroscopy spectrum of a quantum dot prepared in Example 6,

a) CdSe / CdS (-DA) _ex (-MUA-aPEGa) ₅ quantum dots

b) CdSe / CdS (-DA) _ex (-MUA-aPEGa) ₁₀ quantum dots

c) CdSe / CdS (-DA) _ex (-MUA-aPEGa) ₃₀ quantum dots

7 shows a TEM image of a quantum dot prepared in Example 6,

a) CdSe / CdS (-DA) _ex (-MUA-aPEGa) ₅ quantum dots

b) CdSe / CdS (-DA) _ex (-MUA-aPEGa) ₁₀ quantum dots

8 shows infrared spectroscopic spectra of the magnetic nanoparticle starting material and the magnetic nanoparticles prepared in Examples 7 to 11,

a) SPION-OA nanoparticles used as starting material

b) SPION (-OA) _ex (MHA) ₁₀ nanoparticles prepared in Example 7

c) SPION (-OA) _ex (MHA-aPEGa-MTX) ₁₀ nanoparticles prepared in Example 8

d) SPION (-MPA) _ex (MHA-aPEGa-MTX) ₁₀ nanoparticles prepared in Example 9

e) SPION (-Lys) _ex (MHA-aPEGa-MTX) ₁₀ nanoparticles prepared in Example 10

f) SPION (-OA) _ex (MHA-en-FA) ₅ nanoparticles prepared in Example 11

9 shows a transmission electron microscope (TEM) image of the magnetic nanoparticle starting material and the magnetic nanoparticles prepared in Examples 7 to 10,

a) SPION-OA nanoparticles used as starting material

b) SPION (-OA) _ex (MHA) ₁₀ nanoparticles prepared in Example 7

c) SPION (-OA) _ex (MHA-aPEGa-MTX) ₁₀ nanoparticles prepared in Example 8

d) SPION (-MPA) _ex (MHA-aPEGa-MTX) ₁₀ nanoparticles prepared in Example 9

e) SPION (-Lys) _ex (MHA-aPEGa-MTX) ₁₀ nanoparticles prepared in Example 10

10 shows magnetic resonance images (MR) images of experiments in which SPION (-OA) _ex (MHA-en-FA) ₅ magnetic nanoparticles prepared in Example 11 were targeted into KB cells.

*** Explanation of symbols for main parts of drawing ***

10: nanoparticle (core) 12: nanoparticle (shell)

14: surfactant

16: organic ligands wherein at least two hydrophilic groups are linked via a hydrocarbon chain selected from 1 to 7 carbon atoms

18: functional molecule (eg, biocompatible molecule, target oriented molecule, combination or mixture thereof)

20: organic ligand in which a thiol group and a hydrophilic group are linked through a hydrocarbon chain selected from 8 to 20 carbon atoms

A: Hydrophilic Surface Modification Step of Hydrophobic Nanoparticles

B: Functional Molecular Binding Step of Hydrophilic Nanoparticles

C: Partial Hydrophilic Surface Modification Step of Hydrophobic Nanoparticles

D: Functional molecular binding step of the nanoparticles, which includes some hydrophilic moieties but wholly hydrophobic

E: Hydrophilic Surface Modification Step of Functional Nanoparticles Including Some Hydrophilic Part but Showing Hydrophobicity in Overall

Claims (16)

1) The organic ligand is reacted by adding 1 to 30 equivalents of an organic ligand in which a thiol group and a hydrophilic group are linked through a hydrocarbon chain selected from 8 to 20 carbon atoms to a hydrophobic inorganic nanoparticle having a core or a shell / shell structure protected by a surfactant. Chemically bonding a metal-thiolate (MS) covalent bond to the nanoparticle surface while substituting a portion of the surfactant; 2) introduces an irregular surface structure to the surface of the nanoparticles while maintaining individual dispersibility by binding a molecule having a biocompatible, target-oriented or both to the hydrophilic group introduced to the surface of the nanoparticles prepared in step 1) Doing; And 3) converting the remaining surfactant remaining on the surface of the nanoparticles prepared in step 2) with an organic ligand connected with at least two hydrophilic groups through a hydrocarbon chain selected from 1 to 7 carbon atoms to convert them into hydrophilic nanoparticles Including, the method for producing a nano-particles for bio-images. 2. The semiconductor nanostructure according to claim 1, wherein in step 1), the inorganic nanoparticle is composed of one element of zinc, cadmium and lead which are group II elements of the periodic table and one element of sulfur, selenium and tellurium which are group VI elements of the periodic table. Particles, precious metal nanoparticles or iron oxide nanoparticles manufacturing method characterized in that. The method of claim 2, wherein the inorganic nanoparticles are selected from the group consisting of CdSe, ZnS, CdSe / CdS, CdSe / ZnS, Au, Ag, Fe ₂ O _3, and Fe ₃ O ₄ . The method according to claim 1, wherein the organic ligand in step 1) comprises at least one thiol group and at least one hydrophilic group in the molecule, wherein the hydrophilic group is selected from the group consisting of an amine group, a carboxy group, a hydroxyl group and a thiol group Manufacturing method characterized in that. The method of claim 4, wherein the organic ligand is selected from the group consisting of mercaptoundecanoic acid, mercaptododecanoic acid, and mercaptohexadecanoic acid. The method of claim 1, wherein the number of organic ligands that can be chemically bonded to the surface of the nanoparticles by M-S covalent bonding in step 1) is 1 to 30. The group according to claim 1, wherein in step 2), the molecule having biocompatibility, target orientation, or both is selected from the group consisting of an amine group, an aldehyde group, a carboxy group, a hydroxyl group and a thiol group at least at one end thereof. A biocompatible molecule, a target-oriented molecule, a combination or a mixture thereof. The method of claim 1, wherein the organic ligand in step 3) comprises at least two hydrophilic groups in the molecule, wherein the hydrophilic group is selected from the group consisting of an amine group, a carboxy group, a hydroxyl group and a thiol group Way. According to claim 8, wherein the organic ligand is mercaptohexanoic acid (mercaptohexanoic acid), mercaptoacetic acid (mercaptoacetic acid), mercaptopropionic acid (mercaptopropionic acid), dimercaptosuccinic acid, 2-mercaptoethanol (2 mercaptoethanol), 2-aminoethanethiol, lysine (Lysine), arginine (arginine) and aminovaleric acid (aminovaleric acid). The method according to claim 7, wherein the biocompatible molecule has a group selected from an amine group, an aldehyde group and a carboxy group at both chain ends, or at one chain end has one of an amine group, an aldehyde group and a carboxy group and the other chain end. The manufacturing method characterized by having an alkoxy group or a hydroxyl group chosen from 1-7 carbon atoms. The method of claim 10, wherein the biocompatible molecules are polyethylene glycol (PEG), dextran, poly (L-lactide) (PLLA), poly (DL-lactide) (PDLLA), poly-DL-lac Tide / glycolide copolymer (PLGA), chitosan, alginic acid, hyaluronic acid, collagen, heparin and poly (ε-caprolactone). 8. The method according to claim 7, wherein the target-directed molecule has one of an amine group, an aldehyde group, a carboxy group, a hydroxyl group and a thiol group, so that one of an organic ligand or a biocompatible molecule and an amide bond, an ester bond and a thioester bond Manufacturing method characterized in that can be connected by. The method of claim 12, wherein the target oriented molecule is folic acid, methotrexate (MTX), a peptide selective for a specific cell, or an antibody that selectively reacts with a specific antigen. The organic ligand metal is connected to the surface of the surfactant-protected core and the core / shell structured hydrophobic inorganic nanoparticle by a hydrocarbon in which a metal-thiolate-bonded thiol group and a hydrophilic group are selected from 8 to 20 carbon atoms instead of a part of the surfactant. -Thiolate (MS) is chemically bound by covalent bonds, The organic ligand-bonded portion is hydrophilic but exhibits hydrophobicity as a whole. The organic ligand metal is connected to the surface of the surfactant-protected core and the core / shell structured hydrophobic inorganic nanoparticle by a hydrocarbon in which a metal-thiolate-bonded thiol group and a hydrophilic group are selected from 8 to 20 carbon atoms instead of a part of the surfactant. -Thiolate (MS) is chemically bound by covalent bonds, The hydrophilic group of the organic ligand is bonded to a molecule having a biocompatible, target-oriented or both, has an irregular surface structure, The bio-imaging nanoparticles, wherein the portion having the biocompatible, target-oriented or both molecules bound are hydrophilic but generally hydrophobic. The organic ligand metal is connected to the surface of the surfactant-protected core and the core / shell structured hydrophobic inorganic nanoparticle by a hydrocarbon in which a metal-thiolate-bonded thiol group and a hydrophilic group are selected from 8 to 20 carbon atoms instead of a part of the surfactant. -Thiolate (MS) is chemically bound by covalent bonds, The hydrophilic group of the organic ligand is bonded to a molecule having a biocompatible, target-oriented or both, has an irregular surface structure, The remaining surfactant remaining on the surface of the nanoparticles is replaced with an organic ligand connected to at least two hydrophilic groups through a hydrocarbon chain selected from 1 to 7 carbon atoms, so that the entire surface of the nanoparticles is hydrophilic. .

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